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目的:建立液相色谱-串联质谱法(LC-MS/MS法)测定人血浆中氯胺酮及其代谢物去甲基氯胺酮对映异构体浓度,并用于氯胺酮和右氯胺酮临床药动学研究。方法:血浆经固相萃取后,采用PLC-MS/MS测定。色谱柱为酸性糖蛋白结合硅胶柱(100 mm×4mm,5μm),流动相为异丙醇-10 mmol·L~(-1)醋酸胺溶液(氨水调pH至7.6~7.7)(6:94),流速0.5mL·min~(-1),柱温28℃;质谱采用电喷雾离子化(ESI)及多重反应监测(MRM)模式;选择检测离子反应对为m/z 238→m/z207(R-/S-氯胺酮),m/z 224→m/z 207(R-/S-去甲基氯胺酮),m/z 291→m/z 230(内标甲氧氨苄嘧啶)。结果:人血浆中氯胺酮对映异构体的线性范围为5~1000ng·mL~(-1),最低定量限为5ng·mL~(-1);代谢物去甲基氯胺酮对映异构体的线性范围为2.5~500 ng·mL~(-1),最低定量限为2.5 ng·mL~(-1)。本方法专属性良好,氯胺酮对映异构体批内批间精密度均小于5%,去甲基氯胺酮对映异构体的批内批间精密度均小于7%。此检测方法应用于全麻腹腔镜手术受试者静脉注射盐酸氯胺酮注射液或盐酸右氯胺酮后的药代动力学研究中。21例患者单次静脉注射消旋体氯胺酮后,R-氯胺酮和S-氯胺酮半衰期分别为(4.06±1.75)h、(3.33±2.23)h;停药后即刻浓度C_0分别为(3 427±2 866)ng·mL~(-1)、(3331±2797)ng·mL~(-1);24h曲线下面积AUC_(0~24h)分别为(407±91)h·ng·mL~(-1)、(361±87)h·ng·mL~(-1)。代谢物R-N-去甲基氯胺酮和S-N-去甲基氯胺酮半衰期分别为(8.63±2.57)h、(8.73±2.93)h;达峰浓度C_(max)分别为(100±22)ng·mL~(-1)、(91±20)ng·mL~(-1);24h曲线下面积AUC_(0~24h)分别为(557±211)h·ng·mL~(-1)、(515±167)h·ng·mL~(-1)。结论:本方法准确,专属性强,灵敏度高,可满足实际临床研究的需要。
OBJECTIVE: To establish a liquid chromatography-tandem mass spectrometry (LC-MS / MS) method for the determination of ketamine and its metabolite desmethylketamine enantiomers in human plasma and to study the clinical pharmacokinetics of ketamine and dexketamine. Methods: Plasma was extracted by solid-phase extraction and determined by PLC-MS / MS. The column was acid glycoprotein combined with silica gel column (100 mm × 4 mm, 5 μm) and the mobile phase consisted of isopropanol-10 mmol·L -1 ammonium acetate solution (ammonia water adjusted to pH 7.6-7.7) (6:94 ) At a flow rate of 0.5 mL · min ~ (-1) and a column temperature of 28 ℃. The ESI-MS and multiple reaction monitoring (MRM) patterns were used for mass spectrometry. The ion pair was selected as m / z 238 → m / (R- / S-ketamine), m / z 224 → m / z 207 (R- / S-demethylketamine), m / z 291 → m / z 230 (internal standard). Results: The linear range of ketamine enantiomers in human plasma ranged from 5 to 1000 ng · mL -1 with the lowest limit of quantification being 5 ng · mL -1. The metabolites desmethyl ketamine enantiomers The linear range was 2.5 ~ 500 ng · mL ~ (-1) and the lowest limit of quantification was 2.5 ng · mL ~ (-1). The specificity of this method is good, the intra-batch precision of ketamine enantiomers is less than 5%, and the intra-batch precision of demethoxy ketamine enantiomers is less than 7%. This test is used in the pharmacokinetic study of intravenous injection of ketamine hydrochloride or dexketamine hydrochloride in patients undergoing laparoscopic surgery. The half-lives of R-ketamine and S-ketamine were (4.06 ± 1.75) h and (3.33 ± 2.23) h, respectively, after a single intravenous injection of racemic ketamine in 21 patients. The concentrations of C_0 immediately after drug withdrawal were (3 427 ± 2) 866 ng · mL -1 and 3331 ± 2797 ng · mL -1, respectively. The area under the curve of 24h curve was (407 ± 91) h · ng · mL ~ (- 1), (361 ± 87) ng · mL -1. The half-lives of the metabolites RN-demethyl-ketamine and SN-demethyl-ketamine were (8.63 ± 2.57) h and (8.73 ± 2.93) h, respectively; the peak concentrations of Cmax were (100 ± 22) ng · mL ~ (1) and (91 ± 20) ng · mL -1, respectively. The area under 24h curve was (557 ± 211) h · ng · mL -1 167) ng · mL -1. Conclusion: The method is accurate, specific, sensitive and can meet the needs of clinical research.